NeuroPrep™ Medium is unique nutrient-containing formulation optimized for partial enzymatic dissociation of
neuronal tissues such as the NeuroPure™ and NeuroStem™ Primary Rat Neurons. NeuroPrep Medium provides
for rapid and safe enzymatic digestion of primary rodent neuronal tissues when used in conjuction with
NeuroPapain Enzyme (Cat#: NM100200).

Following treatment with NeuroPrep Medium containing NeuroPapain
Enzyme, neuronal tissues are more easily dissociated via mechanical trituration, and the number of viable
dissociated cells can be increased by up to 100%. However, please note that for assays performed within 4 days
of plating, some digestion of surface proteins is inevitable.

NeuroPrep™ Medium

NeuroPrep™ Medium is unique nutrient-containing formulation optimized for partial enzymatic dissociation of
neuronal tissues such as the NeuroPure™ and NeuroStem™ Primary Rat Neurons. NeuroPrep Medium provides
for rapid and safe enzymatic digestion of primary rodent neuronal tissues when used in conjuction with
NeuroPapain Enzyme (Cat#: NM100200).

Following treatment with NeuroPrep Medium containing NeuroPapain
Enzyme, neuronal tissues are more easily dissociated via mechanical trituration, and the number of viable
dissociated cells can be increased by up to 100%. However, please note that for assays performed within 4 days
of plating, some digestion of surface proteins is inevitable.

NeuroPrep medium, 100 ml bottle, NM100100 $69

Note: The following protocol has been validated for use with the NeuroPure Primary Rat Neuronal Cells. Please
refer to the NeuroPure protocols for complete details.

1. Add 5 mg of NeuroPapain Enzyme into 2.5 ml of NeuroPrep Medium. Mix at 37ºC for 15 minutes to completely dissolve the NeuroPapain. Sterilize this solution with a 0.2 μm filter prior to utilizing for tissue digestion. Use within 3 hours for best results.

2. Prior to enzymatic treatment, allow the neuronal tissue to settle for 15 - 30 minutes at 4ºC. Alternatively, place the tube containing the tissue in a 50 ml tube and spin down the cells at 1,100 rpm (200xg) for 1 minute.Transfer the medium from the cell vial to a separate sterile tube while being careful not to remove any loose tissue pieces. Save the medium for trituration following NeuroPapain treatment.

3. Immediately add 2 ml of sterile NeuroPapain solution to the tissue-containing tube, and allow the neuronal tissue to incubate for 30 minutes at 30 ºC. Swirl every two minutes by hand.

4. Following incubation, spin down the cells at 1,100 rpm (200xg) for 1 minute. Remove the NeuroPapain solution, again being careful not to disturb or remove the tissue.

5. Add 1 ml of shipping medium back to the neuronal cells. Save the other 1 ml of shipping medium for Step 4 in
the NeuroPure protocol.